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Callus induction on standard type Cymbidium cultivars
108-110.Views:135Tissue cultured Cymbidium PLBs (protocormlike body) were used as starting material to induce embryogenic callus which could serve as objects of genetic transformation. We obtained callus using two methods. The first method was culturing the PLB segments for one month in liquid MS medium in the presence of 0.5 mg/1 benzyladenine and 0.05 mg/1 naphtylacetic acid followed by cultivation on the same composition solid medium with 0.5 g/l activated charcoal for an additional month. Callus formation was observed on 30% of the explants. The second way was to propagate the PLB segments on solid MS medium supplemented with 1 mg/1 thidiazuron. In these cultures we also observed callus formation on 20% of the explants.
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Down-regulation of ethylene production in carnation (Dianthus Caryphyllus L.) by an apple derived ACC-cDNA
101-104.Views:134Transgenic carnations were produced with an apple derived antisense ACC-synthase cDNA. Transgenic carnation regenerants were potted in glasshouse. All transformed plants showed normal growth and were true-to-type. Ethylene production — measured at full opening stage — lowered by 30-60 %, no plant with 100 % decrease was identified. The vase-life has been observed for 5 years. 38 % of the transformant carnations showed a higher a relative value in days by more than 2 days to 6 days. Twenty six plants were found exhibiting the most marked alterations in the tested trait. In these plants ethylene production decreased by 37-67 %, they have longer vase-life (by 4 days or more). Since the fragrance variety 'Bíbor' was the plant material for genetic modification of vase-life, this trait has been conserved after transformation in spite of the fact that the position of transgene integration cannot be directed.
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Preliminary characterization of the self-incompatibility genotypes of European plum (Prunus domestica L.) cultivars
23-26.Views:233European plum is an important fruit crop with complex, hexaploid genome of unknown origin. The characterization of the selfincompatibility (S) locus of 16 European plum cultivars was carried out using the PaConsI-F primer in combination with the EM-PC1consRD primer for the first intron and the EM-PC2consFD and EM-PC3consRD primers for the second intron amplification. Altogether, 18 different alleles were scored indicating high genetic diversity. These alleles were labelled using alphabetical codes from SA to SS. We identified 5 different alleles in 9 cultivars, 4 alleles in 5 cultivars, while 3 alleles were shown in two of the assayed cultivars. A total of 16 different S-genotypes were assigned, and discrimination of all plum cultivars was successful based on their unique S-genotypes. However, further research is required to reliably identify the S-alleles based on their DNA sequence and clarify complete S-genotypes.
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Microsatellite based identification of grapevine cultivars traditional in Hungary and in the Carpathian Basin
71-73.Views:145Grapevine cultivars and clones traditional in Hungary and in the Carpathian Basin are maintained in the genetic collection of the Corvinus University of Budapest. Mostly ancient varieties and clones were genotyped using microsatellite loci. The investigated 6 loci were sufficient to distinguish all cultivars. Some clones could also be separated based mostly on the variable VVS2 loci. For 17 out of the 31 investigated cultivars this is the first report on characterization of the polymorphism of the allele lengths by microsatellite markers on loci: VVS2, VVMD7, VVMD27, vrZAG62.
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Factors affecting apricot fruit antioxidant capacity and mineral element contents
95-99.Views:225Several epidemiological studies revealed that the consumption of antioxidant compounds and the risk of atherosclerosis, increased blood pressure or cancer are inversely proportional. The individual amounts and relative proportions of macro- and micro elements present in food is also of great consideration since these are involved in a wide range of physiological processes including the influence of the redox homeostasis. The antioxidant power and mineral nutrient content of fruits might be affected by several factors including genotype, ripening stage, year-effects or a wide range of environmental conditions. This study was carried out to survey the antioxidant power and mineral element content in fresh fruits of apricot and analyse some genetic and environmental factors that may have important contribution to the inner content of apricot fruits. In addition, the influencing effect of the extraction procedure used for antioxidant analyses was also tested. Our analyses indicate that a considerable fraction of antioxidant capacity is attributable to the hydrophilic antioxidants. The genetic background has crucial importance in determining apricot fruit antioxidant capacity and mineral nutrient content; however, the growing season and the ripening time of fruits may have also important effects.
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Genotyping Hungarian apricot cultivars for self-(in)compatibility by isoelectric focusing and PCR analysis
69-72.Views:162Self-incompatibility (SI) in flowering plants is a widespread genetic system that promotes out-crossing. In Prunus species the SI is a gametophytic trait, which is controlled by a single multiallelic locus, termed S-locus. S-alleles codify stylar glycoproteins with ribonuclease activity (S-RNases). Our objective was to assess the S-genotype of some Hungarian apricot varieties by isoelectric focusing of stylar RNases as well as by PCR analysis using cherry consensus primers. Consensus primers amplified one or two bands of various sizes. Primers amplifying the 1st intron gained fragments the size of which ranged from 250 to 500 bp; while those amplifying the 2nd intron resulted in fragments of 800-2000 by length. Our data demonstrated that the first intron of the apricot S-RNase gene is shorter than the second one, which coincides with the structure of cherry S-RNase alleles. `Hargrand' (S1S2) and `Harcoe (S1S4) possessed one common S-RNase isoenzyme. Hungarian 'Orias' apricot cultivars showed different bands compared to the previous cultivars, but they shared completely identical patterns confirming that they possess the same S-genotype. 'Bergeron', `Harmat' and 'Korai zamatos' are characterised by an evidently distinct S-RNase pattern. The self-compatible cultivar (`Bergeron') had one allele, which suggests its correspondence to the Sc. Primers for the 2nd intron was unsuccessful in gaining fragments, which indicates that the 2nd intron in the Sc allele is too long to get any amplification. On the basis of our data, identities and differences were revealed in the S-allele constitution of some economically important Hungarian apricot cultivars at protein and DNA levels.
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High-velocity microprojectile mediated DNA delivery into Phaseolus vulgaris callus cells
99-102.Views:123We report the method for the establishment of rapidly growing callus cultures of Phaseolus vulgaris and the conditions required for efficient transformation using high velocity microprojectiles and high level of transient gene expression. Using hypocotyl explant and vertical culture on B5 medium with lmg/1 kinetin and 2 mg/1 2,4-D, we can recommend to get a rapidly growing callus from bean which is a good starting material to introduce foreign DNA into bean cells. The GeneBooster particle delivery system was used for the bombardment of bean callus and the Hgm resistance gene (Hgmr) was used as a selectable marker gene. 25mg/I hygromycin (Hgm) concentration was sufficient to kill the control callus. We used the standard physical factors, the appropriate pressure of N2 gas for the bombardment of the callus tissue, the shooting distance and the size of tungsten particles used as microprojectiles. Selective and nonselective tests were made by transferring the healthy green and white calluses, subcultured for 4 months on selective and nonselective medium. Several Hgm resistant calli had been obtained. Selective pressure was maintained over a period of 10 months.
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Attempting Regeneration from Cultured Cotyledons and Plant Regeneration from Cotyledonary Nodes in Common Bean (Phaseolus vulgaris L.)
57-60.Views:162Dry seeds from two cultivars of common bean (Phaseolus vulgaris L.) were germinated on sterile cotton and sterile deionized distilled water. Cotyledonary node tissue of seedlings were cultured on Murashige and Skoog(MS)-based media supplemented with different combination of N6-benzyl-aminopurine (BAP) and indole-3-acetic acid (IAA), and benzyladenine (BA) and a-naphthaleneacetic acid (NAA). The results revealed that the regeneration percent and the average number of buds and shoots per explant were influenced by the type of explants and exogeneously added hormones. Multiple shoot induction on dry bean cotyledonary node that contain 4-5 mm from cotyledons and hypocotyl on a medium containing full concentration of MS inorganic salts supplemented with 0.5mg/1 BA and 0.1mg/1 NAA was feasible and the method can be applied in transformation experiments.
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Study of different factors of grapevine regeneration systems and genetic transformation
33-36.Views:218The most limitating factor for successful transformation is the absence of high-yielding regeneration protocols. However, the anther-derived embryogenic culture is an optimal technique for genetic transformation and it has been widely applied in many important cultivars, but the necessity of further development of regeneration systems has been proved. We attempted to produce somatic embryos on a wide range of genotypes from various tissues; leaves, petioles, stem segments. We started the examination of grapevine regeneration via organogenesis, succeeded in inducing shoot from the meristematic tissue of the base of bud by testing induction medium contained different concentrations of two types of hormones. To optimize the conditions of the Agrobacterium-mediated transformation, we studied the effectiveness of different Agrobacterium-treatments, the use of antioxidants and the sufficient quantity of kanamycin for selection of transformed cells.
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In vitro regeneration from cotyledons of watermelon
96-98.Views:168Cotyledonary segments of five different genotypes of watermelon were used to induce organogenesis. Five different hormone combinations were applied to enhance the induction of shoot formation on the surface of the segments. The phases of organogenesis were followed with light and scanning electron microscope. Shoots were obtained after four weeks, then the shoots were transferred to hormone free medium for root induction.
This method of regeneration can be applied in transformation experiments. GUS histochemical assay was made to check the expected success of using Agrobacterium for the transformation.
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Examinations of 600-year-old seeds by means of archaeobotanical and genetical methods
79-80.Views:143About 600-year-old plant seeds were discovered in a well of a mediaeval cellar in the course of an excavation in Budapest. After the archaeobotanic purification seed of 16 species were found in large quantities. Seeds preserved in the best state were selected from each group. The existence of endosperm was analysed in these subfossils, which turned to be successful mainly in the case of grapes (Vitis vinifera) and cornels (Cornus mas). Seeds of these two species contained the most endosperm and remains of the embryo. DNA was extracted with the help of DNEasy Plant Mini Kit and analysed by RAPD-PCR method. The amplification of DNA extracted from cornel seeds resulted in detecting a 1500 by fragment, which makes the comparison of these samples with present-day cornels possible.
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Luminescence variations in cucumber (Cucumis sativus L.) leaves derived from different regeneration systems
50-52.Views:159Plants obtained from in vitro culture can show increased susceptibility to environmental stress conditions. In the process of their adaptation to natural conditions it requires monitoring of their physiological state. The methods used to check this phenomenon should estimate quickly and exactly the tolerance to suboptimal environmental factors. Such requirements are satisfied by the methods of measuring chlorophyll luminescence in vivo, e.g. fluorescence induction and delayed luminescence. The objects of our studies were cucumber plants regenerated from cultures of callus and embryogenic cell suspension, as well as the plants obtained from seeds. The plants derived from in vitro cultures displayed a poor physiological condition at the early phase of adaptation characterised by higher susceptibility both to stress caused by increased density of the light flux and low temperature (4 °C) in comparison with the plants obtained from seeds.
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Determination of the cold tolerance off sour cherry cultivars with frost treatments in climatic chamber
49-54.Views:237Nowadays, sour cherry buds can be seriously damaged by spring and winter frosts. Unlike other fruit species threatened by high frost damage, sour cherry cultivars have not been assessed for frost tolerance. The aim of .our survey was to establish the relative cold tolerance of the Hungarian cultivars after treatment in a climatic chamber, and to optimize the methodology formerly elaborated for the frost treatment of apricot. Fourteen cultivars of Hungarian sour cherry (Prunus cerasus) were used in the experiments, which spanned the winters of 2005/2006 and 2006/2007. Our data were used to rank cultivars in two groups according to their levels of cold resistance. We also recommend critical temperatures and treatment times for the testing of sour cherry cultivar resistance to cold in climatic chambers.
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General defense system in the plant kingdom III.
45-54.Views:169Our observations regarding the symptoms not fitting into, significantly differing from the hypersensitive defense system, which we noticed during the judgment of several plant species, symptoms provoked on several million plants have constituted a unified entity. They have provided evidence for the existence of a different plant defense system. We called this so far unknown basic response of plants to biotic effects as general defense system. This system defends them from the attack of numerous microbe species in the environment.
The evolutionary intermediate phase between the general and the specific, the two defense systems is the susceptible host—pathogen relation. The vertical resistance system of plants escaping from the susceptible host—pathogen relation, based on specific hypersensitive reaction also suggested the existence of a more original, general defense system and the susceptible host—pathogen relation developed as a result of the collapse of that system.
The evolutionary relation of the two defense systems is proved by the only recessive inheritance of the older general defense system and in the majority of cases dominant hereditary course of the specific defense system. In our experiences, the modifying genes of the recessive general defense system, in most cases, are behind the specific defense systems, which are known to have monogenic dominant hereditary course and react with hypersensitive tissue destruction. This seemingly striking genetic fact is explained by the following: the general defense system less dependent on environmental effects regulates much faster pathophysiological reaction than the specific resistance genes strongly dependant on environmental effects coding dominant hypersensitive reaction.
The general and specific defense reactions, the processes excluding the microbes attacking plants with compacting of cell growth and tissue destruction, which mean two opposite strategies, building on and regulating each other constitute the entity of resistance to plant disease.
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Review of the molecular background of self-incompatibility in rosaceous fruit trees
7-18.Views:228This review gives a presentation of the gametophytic self-incompatibility system in the roscaeous fruit trees. Studies to discover the pistil (S-ribonucleases) and pollen-part components (F-box molecules) are summarized and models for the self-incompatibility reactions as well as their molecular background are discussed. We describe how mutations within the S-RNase or F-box genes can contribute to the transition from self-incompatibility to the self-compatible phenotype in many fruit tree crops. The current state of the arts is compared to the information obtained in other plant species possessing similar incompatibility system.
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Evaluation of different sources of resistance for breeding powdery mildew resistant grapevine varieties
35-40.Views:180The appearance of resistance to powdery mildew was investigated on seedling progenies of interspecific crossings. Hybrids of American and East-Asian wild species and one Vitis vinifera variety (Janjal kara) were used as source of resistance in the crossings. The resistance of foliage and of berries were tested in the field at the time of vintage. The degree of resistance was ranked into six classes.
In American and Asian hybrid derivatives there were few highly resistant progenies, but in the hybrids of Janjal kara variety more than 90 % of the progenies proved to be highly resistant to powdery mildew. According to the distribution of classes of resistance in the populations the resistance of American and Asian sources are mostly influenced by additive gene affects. The resistance of Janjal kara seemed to be mono or digenic.
No close correlation was found between the powdery mildew resistance of the foliage and that of the berries.
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Transmethylation and the general defense reaction of plants
35-40.Views:176Plant breeding for resistance, namely building specific resistance genes into cultivated plants to ensure resistance against certain pathogen species, is a several-decade-long practice. While looking for purposes of failures appearing during the cultivation of varieties created in this way, a plant feature that ensures non-specific reactions against effects which evoke biotic stress attracted our attention. We named this plant defense form the general defense reaction. The general defense reaction is a fundamental attribute of the plant kingdom, fulfils the role of plant immune system and manifests itself in cell enlargement and cell division. Plants with a high level general defense reaction endure abiotic stresses as well.
In studying the biochemical background of the interaction of the general defense reaction and transmethylation, we found that transmethylation has important role in warding off both biotic and abiotic stresses. According to our observations, plants possessing high level general defense system are suitable for thorough examination of the process and plant physiological role of transmethylation. Biochemical studies also strengthened our observation, which has been taken on the basis of phenotype, that the general defense system can not be ignored during future plant breeding.
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In vitro multiplication and hardening of grapevine plants in aeriated media
15-18.Views:215In vitro cultures have widely been used in horticulture for rapid multiplication of new varieties and clones as well as to produce pathogen-free stock material. To improve efficient hardening and transfer in vitro grown grapevine plants were multiplied by cutting them into single-node internodes with the whole leaf. Microcuttings including the shoot tips were rooted in granulated perlite moisted with tapwater under sterile conditions. After 2-3 weeks the rooted microcuttings were supplied by nutrients and hardened by gradual opening and finally by complete removal of the lids of jars or plastic boxes used for growth. Using this method microcuttings of Vitis vinifera cvs. „Chardonnay", „Cabernet franc", „Riesling" and „Sauvignon blanc" and the rootstock varieties Vitis riparia x Vitis cinerea cv. „Barrier" and Vitis berlandieri x Vitis rupestris cv. „Richter 110" formed new roots and shoots and 100% of the tested plants survived the acclimatization procedure. Similar results were obtained when perlite was replaced with rockwool-, or pit-pot blocks. This method may highly increase the efficiency of producing pathogen-free propagating material and new transgenic lines.
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Resistance Gene Analogs (RGA) as a tool in fruit tree's breeding
7-15.Views:187Breeding for pest and disease resistance comes as a major objective behind the fruit traits. To increase the effectiveness of fruit resistance breeding application of the Marker Assisted Selection ( MAS) is advantageous. For generating molecular markers which enable the following of interesting traits basically two methods are available: targeted marker design based on conservative region of already known Resistance ( R) gene sequences or randomly generated markers. The creation and the application of these homology based markers are the object of this review in the main temperate zone fruit species.
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The effects of ACS (1-aminocyclopropane-l-carboxylate synthase) gene down regulation on ethylene production and fruit softening in transgenic apple
65-70.Views:126A detailed examination of the production of ethylene and other ripening parameters during storage period has been undertaken in transgenic apple fruits, where the ethylene biosynthesis was inhibited by antisense ACS (l-aminocyclopropane-l-carboxylate synthase) gene. Data indicate down regulation of ethylene production, softening and spoilage in some transgenic lines. In some cases ethylene production was inhibited for over 90 percent, considerable reduction of softening and spoilage was observed probably due to the reduced activity of cell wall degradable enzymes. ACS activity was also monitored during ripening. The fruits of the best transgenic lines could be stored for minimum 4-5 months longer under 5 °C cold room storage conditions and one month longer at normal room temperature. This molecular approach can provide an alternative way to replace the commonly used and costly atmospheric storage of fruits.
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Analyses of Hungarian sour cherry germplasm with simple sequence repeat markers
27-31.Views:311Twenty-four sour cherry cultivars (genotypes), belonging to four cultivar groups were fingerprinted using microsatellite markers. All genotypes have been arisen from the Carpathian basin, which could be secondary gene centre of sour cherry, since its progenitor species, ground cherry and sweet cherry overlap here. Five SSR primer pairs, earlier used for fingerprinting Turkish sour cherry germplasm were tested. None of the five primer pairs showed any polymorphism within the cultivar groups. The primer pairs were able to distinguish between the cultivar groups. The Oblacsinszka and the Cigánymeggy cultivar groups were the most difficult to separate, while the Pándy cultivar group was the most distinguishable.
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Review of nutritional value and putative health-effects of quince (Cydonia oblonga Mill.) fruit
29-32.Views:499Quince (Cydonia oblonga Mill.) has been long regarded to be a health-promoting fruit. Although it is mainly consumed in processed form, its relatively heat-stable polyphenolics were confirmed to be responsible for most of the beneficial effects. This review aims to show basic nutritional information on quince fruit such as carbohydrate, fibre, pectin and mineral element contents. In addition, vitamin and polyphenolic contents and composition as well as the physiological effects of quince consumption were also surveyed. Information on presumable protective effects against several diseases including infl ammation, atopic dermatitis, ulcers and cancer is summarized. Potential antibacterial effects of quince polyphenolics were also considered. Polyphenolics are supposed to be responsible for the major part of benefi cial health-effects, and phenolic compounds predominantly accumulate in peel. There exists a considerable extent of genetic variation in phytochemical composition among cultivars, which might be exploited in designing future breeding programs for quince improvement and opening new ways for health-related uses.
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The inheritance and durability of scab resistance in apple progenies
39-46.Views:144In order to select the appropriate parent cultivars and maintain the durability of resistance, it is important to clarify the mechanisms of inheritance of scab resistance depending on the parents. It has been known that the progeny segregation ratios based on scab-resistance do not depend only on the genotype of the resistance locus but also on the genetical makeup of the donor and recipient parents as well as on the susceptible parent.
The aim of this study has been to demonstrate what factors in the Vf, Vr and VA scab-resistant cultivars — combined with susceptible and resistant parents — affect the inheritance and durability of resistance in seedlings in their first 4 year's growing four years' growth. After inoculating apple seedlings sown in 2001 with the suspension of Venturia inaequalis (Cke.) Wint. in the greenhouse, we studied the segregation ratios of the progenies into reaction classes. Seedlings showing resistance in the greenhouse were also evaluated for scab-resistance after they had been moved to the field and had naturally been infected with the pathogen in 2002 and 2004.
The majority of our results obtained in the greenhouse test, similarly to earlier experiences, have not justified monogenic inheritance at the phenotypic level. The effect of susceptible parent cultivars on the segregation ratio of progenies have become apparent again. The high infection rate of seedlings in the field trials, which had previously exhibited varying degrees of resistance in the greenhouse test, has raised concern. Our data has raised further doubts, concerning the durability of Vf resistance in Hungary. It is assumed that the composition of natural field populations of Venturia inaequalis in Szigetcsép has changed. The complexity of Vf resistance has been confirmed. The high infection rate in the progenies derived from Vf resistant cultivars draws the attention to the importance of utilizing additional sources of resistance.
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Old Hungarian grapevine cultivars and their relations characterized with microsatellite markers
27-31.Views:226Thirty-one old Hungarian grapevine (Vitis vinifera L.) cultivars were investigated on 7 microsatellite loci to characterize them, to separate the cultivars from synonym names, and to confirm parent-offspring connections. Conculta (group of cultivars or bud sports) members, such as `Goher' and Tajor' representatives, were studied to find a suitable locus for the separation. Synonyms, conculta members, subcultivars and clones of Turmine, which was the most important cultivar of Tokaj, were also analyzed to separate the members of the different taxonomic levels. Pedigree of 'Kiralyleanyka' was examined to find the missing ancestor, because the parent-offspring connection between the natural hybrid and `Koverszolo' is questionable.
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A critical evaluation of methods used for S-genotyping: from trees to DNA level
19-29.Views:198Fruit setting behaviour of fruit trees remains to be in the focus of plant breeders and growers. Realizing that most species (cherry, apple, pear etc.) are self-incompatible and certain cultivars are cross-incompatible, mutual fertility properties and their reliable determination are of great interest. This review gives a comprehensive description of all known S-genotyping procedures, i.e. the classical fruit set analysis after open field test crosses; pollen tube growth monitoring with fluorescent microscopy; stylar ribonuclease electrophoresis (using different types of isoelectric focusing and 2-dimension polyacrilamide gel electrophoresis); as well as the most recent polymerase chain reaction based DNA-level analyses and DNA sequencing. The review presented not only gives a compilation of the bases of the methods described but also provides a critical evaluation and a comparative characterization of their applicability.